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Menstrual blood preserving fluid and application thereof and separation culture method of menstrual blood-source endometrium stem cells

A technology for separating, culturing and preserving liquid, applied in the direction of non-embryo pluripotent stem cells, application, animal cells, etc., can solve the problems of small number of stem cells, contaminated menstrual blood, low purity, etc., to ensure purity and vitality, avoid menstrual blood pollution, The effect of reducing pollution rate

Active Publication Date: 2016-05-04
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, for the isolation and culture of endometrial stem cells, most of them are directly obtained, which causes great harm to patients, and at the same time, the source is suffering and is subject to ethical restrictions. Although there is a small amount of research on menstrual blood-derived stem cells, there is no simple and convenient method. Collection of menstrual blood leads to a higher probability of menstrual blood contamination, fewer stem cells obtained, lower purity, lower differentiation potential, higher cost and complicated process, etc.

Method used

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  • Menstrual blood preserving fluid and application thereof and separation culture method of menstrual blood-source endometrium stem cells
  • Menstrual blood preserving fluid and application thereof and separation culture method of menstrual blood-source endometrium stem cells
  • Menstrual blood preserving fluid and application thereof and separation culture method of menstrual blood-source endometrium stem cells

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Experimental program
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Embodiment approach

[0044] Concrete embodiments of the present invention are as follows:

[0045] 1. Preparation of menstrual blood collection kit:

[0046] Including a sterile specimen preservation bottle (containing 5-15ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual.

[0047] The menstrual blood preservation solution contains (2-5)×penicillin, (2-5)×streptomycin, 30-50μg / ml kanamycin, 30-50μg / ml sodium citrate, 100U / ml heparin sodium Phosphate buffer.

[0048] 2. Collection of menstrual blood

[0049] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0050] 3. Sterility test:

[0051] Under sterile conditions, take 0.5-1ml of a sample an...

Embodiment 1

[0063] 1. Preparation of menstrual blood collection kit:

[0064] Including a sterile specimen preservation bottle (containing 5ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual.

[0065] The menstrual blood preservation solution is a phosphate buffer solution containing 2×penicillin, 5×streptomycin, 40 μg / ml kanamycin, 50 μg / ml sodium citrate, and 100 U / ml heparin sodium.

[0066] 2. Collection of menstrual blood

[0067] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0068] 3. Sterility testing:

[0069] Under sterile conditions, take 0.5-1ml of a sample and use a rapid microbial detector to detect whether there is bact...

Embodiment 2

[0080] 1. Preparation of menstrual blood collection kit:

[0081] Including a sterile specimen preservation bottle (containing 15ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual. The menstrual blood preservation solution is a phosphate buffer solution containing 3×penicillin, 4×streptomycin, 50 μg / ml kanamycin, 30 μg / ml sodium citrate, and 100 U / ml heparin sodium.

[0082] 2. Collection of menstrual blood

[0083] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0084] 3. Sterility testing:

[0085] Under sterile conditions, take 0.5-1ml of a sample and use a rapid microbial detector to detect whether there is bacterial co...

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Abstract

The invention relates to the field of biotechnology and particularly relates to a menstrual blood preserving fluid and an application thereof and a separation culture method of menstrual blood-source endometrium stem cells, wherein the menstrual blood is the source of the menstrual blood-source endometrium stem cells. The menstrual blood of women is used as the source for stem cell collection, so that the waste is utilized and turned into wealth while the source is abundant; the number of endometrium stem cells in the menstrual blood is 30 times that in bone marrow, and the active ingredients are plentiful; and the menstrual blood is preserved by the menstrual blood preserving fluid so as to prevent the menstrual blood from being contaminated by the outside in a process from collection to separation. In the invention, the menstrual blood is preserved by the menstrual blood preserving fluid with a unique formula, and the contamination rate of the menstrual blood is remarkably reduced while the purity and activity of the obtained menstrual blood-source endometrium stem cells are guaranteed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a menstrual blood preservation solution and its application, and a method for isolating and culturing endometrial stem cells derived from menstrual blood. Background technique [0002] The uterus is a tissue with high regenerative and differentiation capabilities. In each menstrual cycle, the endometrium of a woman can grow from about 0.5mm to about 5-7mm. A woman can experience 400 such cycles in her lifetime. Studies have shown that the endometrium contains stem cells with high differentiation potential, proliferative ability and low immunogenicity. However, its direct acquisition methods, including hysterectomy, early pregnancy hysterectomy, and curettage, are all severely invasive to the donor. [0003] In recent years, Japanese scientists have discovered stem cells with multi-directional differentiation potential in women's menstrual blood, which can be used to repair damaged m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/074
CPCA01N1/021C12N5/0682C12N2500/90
Inventor 陈海佳王一飞葛啸虎马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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